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Annals of Clinical & Laboratory Science 37:152-157 (2007)
© 2007 Association of Clinical Scientists

Simple Immunohistochemical Staining Method Using Large Sized Gold Colloid Conjugated Secondary Antibody

Eun Jung Lee1, Yonggoo Kim1, Jihyang Lim1, Myungshin Kim1, Chang Suk Kang2, Jae Hoon Lee2, Rak Sun Mok2, Young Nam Park2, Ha-Young Choi1 and Kyungja Han1
1 Department of Laboratory Medicine, Hospital Pathology, Catholic University Medical College, Seoul; 2 Research and Development Center, YD Diagnostics Corp., Yongin Kyunggi-Do, Korea

Address correspondence to Kyungja Han, M.D., Department of Laboratory Medicine, Catholic University Medical College, St. Mary’s Hospital, Youngdeungpo-gu, Youidodong 62, Seoul, Korea [South] 150-713; tel 82 2 3779 1297; fax 82 2 783 6648; e-mail hankja{at}catholic.ac.kr.

To test the feasibility of semiquantitative immunohistochemical staining (IHC) with large sized gold conjugated secondary antibody (gold-2°Ab), we used beads covered with a known amount of primary antibody and a secondary antibody conjugated with gold colloid particles (20 and 40 nm in diameter), and we compared the results to those obtained by enzyme IHC. Beads coated with 6 graded amounts of mouse IgG molecules showed 6 levels of color intensity. The graded color intensities could readily be distinguished. The color developed as soon as we added gold-2°Ab, and the intensities were stable for 1 wk. Enzyme IHC using identical beads showed dregs of pigment after incubation in DAB for 5 min. The large sized gold-2°Ab showed strong signals on cell surfaces; application of the large sized gold-2°Ab to paraffin-embedded tissue sections was also feasible. The color was bright red and was easier to differentiate from hemosiderin pigment than the color developed by enzyme IHC. In conclusion, gold IHC with large sized gold-2°Ab is superior to enzyme IHC for quantification of antigens via IHC. Gold IHC is especially recommended for tissues with many macrophages, such as bone marrow and spleen.

Keywords: gold colloid particles, immunohistochemistry, gold conjugated secondary antibody




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Y. Moon, G. Park, K. Han, C.-S. Kang, and W. Lee
Mouse Spleen Tissue as a Staining Intensity Reference for Immunohistochemistry
Ann. Clin. Lab. Sci., January 1, 2008; 38(3): 215 - 220.
[Abstract] [Full Text] [PDF]


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